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Journal article
Transcriptional initiation is controlled by upstream GC-box interactions in a TATAA-less promoter
Molecular and cellular biology, v 10(12), pp 6632-6641
01 Dec 1990
PMID: 2247077
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Abstract
Numerous genes contain TATAA-less promoters, and the control of transcriptional initiation in this important promoter class is not understood. We have determined that protein-DNA interactions at three of the four proximal GC box sequence elements in one such promoter, that of the hamster dihydrofolate reductase gene, control initiation and relative use of the major and minor start sites. Our results indicate that although the GC boxes are apparently equivalent with respect to factor binding, they are not equivalent with respect to function. At least two properly positioned GC boxes were required for initiation of transcription. Abolishment of DNA-protein interaction by site-specific mutation of the most proximal GC box (box I) resulted in a fivefold decrease in transcription from the major initiation site and a threefold increase in heterogeneous transcripts initiating from the vicinity of the minor start site in vitro and in vivo. Mutations that separately abolished interactions at GC boxes II and III while leaving GC box I intact affected the relative utilization of both the major and minor initiation sites as well as transcriptional efficiency of the promoter template in in vitro transcription and transient expression assays. Interaction at GC box IV when the three proximal boxes were in a wild-type configuration had no effect on transcription of the dihydrofolate reductase gene promoter. Thus, GC box interactions not only are required for efficient transcription but also regulate start site utilization in this TATAA-less promoter.
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Details
- Title
- Transcriptional initiation is controlled by upstream GC-box interactions in a TATAA-less promoter
- Creators
- M C Blake - University of North Carolina at Chapel HillR C Jambou - University of North Carolina at Chapel HillA G Swick - University of North Carolina at Chapel HillJ W Kahn - University of North Carolina at Chapel HillJ C Azizkhan - University of North Carolina at Chapel Hill
- Publication Details
- Molecular and cellular biology, v 10(12), pp 6632-6641
- Publisher
- American Society for Microbiology (ASM)
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Biochemistry and Molecular Biology
- Web of Science ID
- WOS:A1990EJ60200057
- Other Identifier
- 991020200886404721
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- Web of Science research areas
- Biochemistry & Molecular Biology
- Cell Biology